Simultaneous determination of intracellular concentrations of tenofovir, emtricitabine, and dolutegravir in human brain microvascular endothelial cells using liquid chromatography-tandem mass spectrometry (LC-MS/MS)

Anal Chim Acta. 2019 May 16:1056:79-87. doi: 10.1016/j.aca.2019.01.015. Epub 2019 Jan 18.

Abstract

Combination antiretroviral therapy (cART) regimens are recommended for HIV patients to better achieve and maintain plasma viral suppression. Despite adequate plasma viral suppression, HIV persists inside the brain, which is, in part thought to result from poor brain penetration of antiretroviral drugs. In this study, a simple and ultra-sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of tenofovir, emtricitabine, and dolutegravir in cell lysates of an immortalized human brain microvascular endothelial cell line (hCMEC/D3) was developed and validated. Analytes were separated on a reverse phase C18 column using water and 0.1% formic acid in acetonitrile as mobile phases. The analytes were detected using positive electrospray ionization mode with multiple reaction monitoring (MRM). The assay was linear in the concentration range of 0.1-100 ng mL-1 for all analytes. Intra- and inter-assay precision and accuracy were within ±13.33% and ±10.53%, respectively. This approach described herein was used to determine the intracellular accumulation of tenofovir, emtricitabine, dolutegravir simultaneously in hCMEC/D3 cells samples.

Keywords: Antiretroviral drugs; Dolutegravir; Emtricitabine; HIV; Mass spectrometry; Tenofovir.

MeSH terms

  • Analytic Sample Preparation Methods
  • Anti-HIV Agents / analysis*
  • Anti-HIV Agents / isolation & purification
  • Brain / cytology*
  • Cell Line
  • Chromatography, Liquid / methods*
  • Emtricitabine / analysis
  • Emtricitabine / isolation & purification
  • Endothelial Cells / cytology*
  • Heterocyclic Compounds, 3-Ring / analysis
  • Heterocyclic Compounds, 3-Ring / isolation & purification
  • Humans
  • Intracellular Space / chemistry*
  • Linear Models
  • Oxazines
  • Piperazines
  • Pyridones
  • Reproducibility of Results
  • Tandem Mass Spectrometry / methods*
  • Tenofovir / analysis
  • Tenofovir / isolation & purification

Substances

  • Anti-HIV Agents
  • Heterocyclic Compounds, 3-Ring
  • Oxazines
  • Piperazines
  • Pyridones
  • Tenofovir
  • dolutegravir
  • Emtricitabine